[1]王栋,潘浩,朱杭,等.强筋壮骨祛风合剂对髓核致炎大鼠背根神经节中3型酸敏感离子通道的影响[J].中医正骨,2017,29(01):18-22,26.
　WANG Dong,PAN Hao,ZHU Hang,et al.Effect of Qiangjin Zhuanggu Qufeng Heji on acid-sensing ion channel 3 of dorsal root ganglia in rats with inflammation caused by nucleus pulposus[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):18-22,26.
点击复制

强筋壮骨祛风合剂对髓核致炎大鼠背根神经节中3型酸敏感离子通道的影响()
分享到：

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期数:
2017年01期
页码:
18-22,26
栏目:
基础研究
出版日期:
2017-01-20

文章信息/Info

Title:
Effect of Qiangjin Zhuanggu Qufeng Heji on acid-sensing ion channel 3 of dorsal root ganglia in rats with inflammation caused by nucleus pulposus
作者:
王栋; 潘浩; 朱杭; 诸力; 何永江; 王坚; 贾高永
浙江省杭州市中医院,浙江 杭州 310007
Author(s):
WANG Dong; PAN Hao; ZHU Hang; ZHU Li; HE Yongjiang; WANG Jian; JIA Gaoyong
Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310007,Zhejiang,China
关键词:
腰痛;  强筋壮骨祛风合剂;  3型酸敏感离子通道;  神经节; 脊;  大鼠; Sprague-Dawley;  动物实验
Keywords:
low back pain;  Qiangjin Zhuanggu Qufeng Heji;  acid-sensing ion channel 3;  ganglia; spinal;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:观察强筋壮骨祛风合剂对髓核致炎大鼠背根神经节中3型酸敏感离子通道(acid-sensing ion channel 3,ASIC3)的影响。方法:选取40只清洁级成年雄性SD大鼠,随机分为空白组、假手术组、模型组、阿米洛利组和中药组,每组8只。除空白组外,其余各组大鼠均手术暴露L5背根神经节。假手术组暴露L5背根神经节后直接缝合切口; 模型组和中药组取出大鼠尾部的髓核组织置于暴露的L5背根神经节上,缝合切口; 阿米洛利组将取自大鼠尾部的髓核组织置于暴露的L5背根神经节上,并在暴露的L5背根神经节周围滴入含0.1 mg盐酸的阿米洛利药液1 mL,缝合切口。自造模后第1天开始,中药组按5 mL·kg-1以强筋壮骨祛风合剂灌胃,每天2次,连续30 d; 空白组、假手术组、模型组不进行药物干预。分别于造模前1 d及造模后3、5、9、15、23、29 d,采用Up-Down法测定大鼠左后足50%机械刺激缩足阈值(50% paw withdrawal threshold,50%PWT)。中药组药物干预结束后,处死所有大鼠,取出L5背根神经节分成2份,一份采用免疫组织化学法计算ASIC3阳性面积,另一份采用Western Blot技术检测ASIC3蛋白表达量。结果:造模后3 d内,模型组、阿米洛利组和中药组均有部分大鼠出现左后足外翻以及足趾背伸现象。造模前后不同时间50%PWT的差异有统计学意义,即存在时间效应(F=50.132,P=0.000)。5组50%PWT总体上比较,组间差异有统计学意义,即存在分组效应(F=288.219,P=0.000)。除造模前1 d时外(F=0.332,P=0.854),造模后各时点5组大鼠50%PWT比较,组间差异均有统计学意义(F=288.015,P=0.000; F=308.588,P=0.000; F=374.945,P=0.000; F=560.713,P=0.000; F=343.043,P=0.000; F=459.914,P=0.000); 造模后各时点模型组的50%PWT均低于空白组、假手术组、阿米洛利组和中药组(P=0.010,P=0.011,P=0.001,P=0.001; P=0.023,P=0.022,P=0.000,P=0.004; P=0.013,P=0.013,P=0.003,P=0.001; P=0.012,P=0.002,P=0.002,P=0.002; P=0.004,P=0.000,P=0.000,P=0.000; P=0.003,P=0.005,P=0.003,P=0.013); 造模后各时点空白组和假手术组50%PWT比较,差异均无统计学意义(P=0.640,P=0.710,P=0.590,P=0.480,P=0.330,P=0.390); 造模后各时点阿米洛利组50%PWT均高于中药组(P=0.021,P=0.011,P=0.013,P=0.002,P=0.001,P=0.005)。时间因素与分组因素存在交互效应(F=2.358,P=0.002)。5组大鼠背根神经节中ASIC3阳性面积和ASIC3蛋白表达量比较,组间差异均有统计学意义(F=205.890,P=0.030; F=101.740,P=0.001); 空白组、假手术组、阿米洛利组和中药组背根神经节中ASIC3阳性面积和ASIC3蛋白表达量均小于模型组(P=0.004,P=0.003,P=0.001,P=0.007; P=0.000,P=0.000,P=0.000,P=0.000); 阿米洛利组背根神经节中ASIC3阳性面积和ASIC3蛋白表达量均大于中药组(P=0.001; P=0.000)。结论:强筋壮骨祛风合剂可以降低髓核致炎大鼠病变的背根神经节中ASIC3的水平,提高大鼠的疼痛阈值,但其作用效果不及盐酸阿米洛利。
Abstract:
Objective:To observe the effect of Qiangjin Zhuanggu Qufeng Heji(强筋壮骨祛风合剂,QJZGQFHJ)on acid-sensing ion channel 3(ASIC3)of dorsal root ganglia(DRG)in rats with inflammation caused by nucleus pulposus.Methods:Forty clean adult male SD rats were selected and randomly divided into blank group,sham-operated group,model group,Amiloride group and TCD group,8 cases in each group.The surgeries were performed on rats in sham-operated group,model group,Amiloride group and TCD group to expose L5 DRG.The surgical incisions were sutured in sham-operated group right after the L5 DRG were exposed.The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in model group and TCD group,and then the surgical incisions were sutured.The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in Amiloride group,and Amiloride solution(1 ml),containing hydrochloric acid(0.1 mg),was dropped arround the denudate L5 DRG,then the surgical incisions were sutured.Since the 1st day after modeling,the rats in TCD group were intragastric administrated with QJZGQFHJ(5ml/kg),twice a day for 30 consecutive days.No drug interventions were performed on rats in blank group,sham-operated group and model group.The 50% paw withdrawal threshold(50%PWT)of left posterior feet were measured by using Up-Down methods at 1 day before modeling and at 3,5,9,15,23 and 29 days after modeing respectively.After the end of drug interventions,all rats in TCD group were executed and the L5 DRGs were fetched out and were divide into 2 parts.One of them was used for calculating the ASIC3 positive areas by using immunohistochemical method,and the other potion was used for detecting the ASIC3 protein expression by using Western Blot assay.Results:Left posterior foot eversion and toe dorsal extension were found in some rats of model group,Amiloride group and TCD group within 3 days after modeling.There was statistical difference in 50% PWT between different timepoints before and after modeling,in other words,there was time effect(F=50.132,P=0.000).There was statistical difference in 50% PWT between the 5 groups in general,in other words,there was group effect(F=288.219,P=0.000).Except at 1 day before modeling(F=0.332,P=0.854),there was statistical difference in 50% PWT between the 5 groups(F=288.015,P=0.000; F=308.588,P=0.000; F=374.945,P=0.000; F=560.713,P=0.000; F=343.043,P=0.000; F=459.914,P=0.000).At different timepoints after modeling,the 50% PWT were lower in model group compared to blank group,sham-operated group,Amiloride group and TCD group(P=0.010,P=0.011,P=0.001,P=0.001; P=0.023,P=0.022,P=0.000,P=0.004; P=0.013,P=0.013,P=0.003,P=0.001; P=0.012,P=0.002,P=0.002,P=0.002; P=0.004,P=0.000,P=0.000,P=0.000; P=0.003,P=0.005,P=0.003,P=0.013).There was no statistical difference in 50%PWT between blank group and sham-operated group(P=0.640,P=0.710,P=0.590,P=0.480,P=0.330,P=0.390)and the 50% PWT were higher in Amiloride group compared to TCD group(P=0.021,P=0.011,P=0.013,P=0.002,P=0.001,P=0.005)at different timepoints after modeling.There was interaction between time factor and group factor(F=2.358,P=0.002).There was statistical difference in ASIC3 positive areas and ASIC3 protein expression in DRG between the 5 groups(F=205.890,P=0.030; F=101.740,P=0.001).ASIC3 positive areas and ASIC3 protein expression were less in blank group,sham-operated group,Amiloride group and TCD group compared to model group(P=0.004,P=0.003,P=0.001,P=0.007; P=0.000,P=0.000,P=0.000,P=0.000),and were greater in Amiloride group compared to TCD group(P=0.001; P=0.000).Conclusion:QJZGQFHJ can reduce the ASIC3 level in diseased DRG and increase the pain threshold in rats with inflammation caused by nucleus pulposus,while it is inferior to Amiloride in effectiveness.

参考文献/References:

[1] 舒剑臣,唐小穗,赵京元.腰椎间盘退变引起腰腿痛的相关化学因素研究进展[J].中华临床医师杂志:电子版,2012,6(6):1534-1537.
[2] HUMZAH MD,SOAMES RW.Human intervertebral disc:structure and function[J].Anat Rec,1988,220(4):337-356.
[3] SLUKA KA,RASMUSSEN LA,EDGAR MM,et al.Acid-Sensing ion Channel 3 deficiency increases inflammation but decreases pain behavior in murine arthritis[J].Arthritis Rheum,2013,65(5):1194-1202.
[4] WEMMIE JA,TAUGHER RJ,KREPLE CJ.Acid-sensing ion channels in pain and disease[J].Nat Rev Neurosci,2013,14(7):461-471.
[5] LI WG,YU Y,ZHANG ZD,et al.ASIC3 channels integrate agmatine and multiple inflammatory signals through the nonproton ligand sensing domain[J].Mol Pain,2010,6:88.
[6] 贺秋兰,魏明.酸敏感离子通道ASIC3与炎性痛觉敏感关系[J].实用医学杂志,2010,26(15):2843-2845.
[7] CHAPLAN SR,BACH FW,POGREL JW,et al.Quantitative assessment of tactile allodynia in the rat paw[J]. J Neurosci Methods,1994,53(1):55-63.
[8] FREBURGER JK,HOLMES GM,AGANS RP,et al.The rising prevalence of chronic low back pain[J].Arch Intern Med,2009,169(3):251-258.
[9] OHTORI S,INOUE G,KOSHI T,et al.Up-regulation of acid-sensing ion Channel 3 in dorsal root ganglion neurons following application of nucleus pulposus on nerve root in rats[J].Spine(Phila Pa 1976),2006,31(18):2048-2052.
[10] 杨宇,何睿,张宇宁,等.酸敏感离子通道与炎性痛中枢敏化机制的研究[J].中国现代药物应用,2014,09(9):228-228.
[11] 郭婕,张前德.补肾方骨青颗粒对佐剂性关节炎大鼠的干预和影响软骨表达酸敏感离子通道3的实验研究[J].南京医科大学学报(自然科学版),2012,32(6):773-778.
[12] 杨廷智,罗兵,杨明庭.加味独活寄生汤配合手法治疗腰椎间盘突出症100例[J].陕西中医,2012,33(12):1606-1607.
[13] 李国强.身痛逐瘀汤加味合电针、牵引治疗腰椎间盘突出症93例临床观察[J].四川中医,2009,26(12):117-118.
[14] 王久瑞.补肾化瘀法治疗腰椎间盘突出症临床观察[J].中医学报,2013,28(8):1239-1240.

相似文献/References:

[1]宁兴明,伍亮,王廷,等.五禽戏配合核心肌力训练治疗非特异性腰痛的临床研究[J].中医正骨,2015,27(11):25.
　NING Xingming,WU Liang,WANG Ting,et al.Clinical study on five mimic-animal boxing combined with core muscular strength exercise for the treatment of nonspecific low back pain[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(01):25.
[2]金立昆,齐越峰,唐可,等.中药离子导入联合腰背部功能锻炼治疗 非特异性腰痛的临床研究[J].中医正骨,2016,28(01):20.
　JIN Likun,QI Yuefeng,TANG Ke,et al.Clinical study on iontophoresis of traditional Chinese medicine combined with lower back functional exercise for treatment of non-specific low back pain[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(01):20.
[3]杨勇,王雷生,张娟.经椎关节突关节温针治疗腰椎关节突关节源性腰痛[J].中医正骨,2016,28(02):67.
[4]袁启令,刘亮,马江涛,等.针刺治疗慢性非特异性腰痛的临床研究[J].中医正骨,2016,28(06):12.
　YUAN Qiling,LIU Liang,MA Jiangtao,et al.A clinical study of acupuncture therapy for treatment of chronic nonspecific low back pain[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(01):12.
[5]刘艳,谢新立,杜彪,等.SPECT全身骨显像在儿童和青少年腰痛诊断中的价值[J].中医正骨,2017,29(04):42.
[6]吴婷婷,杨京辉,汪亚群,等.针刺联合太极拳训练治疗慢性非特异性腰痛的临床研究[J].中医正骨,2017,29(06):32.
　WU Tingting,YANG Jinghui,WANG Yaqun,et al.Clinical study on acupuncture therapy combined with Taijiquan(太极拳)exercise for treatment of chronic nonspecific low back pain[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):32.
[7]熊国星,王鑫,程元辉,等.动推疗法与单纯推拿疗法治疗慢性非特异性腰痛的近期疗效对比研究[J].中医正骨,2017,29(11):33.
　XIONG Guoxing,WANG Xin,CHENG Yuanhui,et al.A comparative study of short-term clinical curative effect of massage during exercise versus monotherapy of TUINA for treatment of chronic nonspecific low back pain[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):33.
[8]李展新,刘建航,李锦威,等.韦氏脊柱整治手法结合核心肌群训练治疗非职业高尔夫运动者非特异性腰痛[J].中医正骨,2018,30(04):13.
　LI Zhanxin,LIU Jianhang,LI Jinwei,et al.Wei's spinal remediation manipulation combined with core muscle training for treatment of nonspecific low back pain in non-professional golfers[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(01):13.
[9]王雷生,杨勇.经膀胱经腧穴透刺关节突关节温针灸治疗腰椎关节突关节源性腰痛的临床研究[J].中医正骨,2018,30(05):6.
　WANG Leisheng,YANG Yong.A clinical study of acupuncture at zygapophysial joint with warmed needle that reaches there by penetrating acupoint of bladder meridian for treatment of low back pain originated from lumbar zygapophysial joints[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(01):6.
[10]叶晓宇,徐卫星,王润民.非手术与微创手术治疗椎间盘源性腰痛的研究进展[J].中医正骨,2018,30(05):40.

备注/Memo

备注/Memo:
基金项目:浙江省中医药科技计划项目(2015ZA157); 浙江省杭州市科技计划重点专病专科项目(20150733Q60)
通讯作者:潘浩 E-mail:Harper1966@163.com
更新日期/Last Update: 2017-01-20